Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Minimal Fuzzy Memberships and Rules Using Hierarchical Genetic Algorithms

Abstract—A new scheme to obtain optimal fuzzy subsets and rules is proposed. The method is derived from the use of genetic algorithms, where the genes of the chromosome are classified into two different types. These genes can be arranged in a hierarchical form, where one type of genes controls the other type of genes. The effectiveness of this genetic formulation enables the fuzzy subsets and rules to be optimally reduced and, yet, the system performance is well maintained. In this paper, the details of formulation of the genetic structure are given. The required procedures for coding the fuzzy membership function and rules into the chromosome are also described. To justify this approach to fuzzy logic design, the proposed scheme is applied to control a constant water pressure pumping system. The obtained results, as well as the associated final fuzzy subsets, are included in this paper. Because of its simplicity, the method could lead to a potentially low-cost fuzzy logic implementation. Index Terms—DNA, fuzzy control, genetic algorithms. I